促红细胞生成素衍生肽的制备和鉴定及对大鼠肾缺血再灌注损伤的保护作用

Preperation and Indentification of A Helix B-surface Peptide and Its Protective Role on Renal Ischemia Reperfusion Injury

目的：制备并鉴定促红细胞生成素衍生肽（HBSP）并探讨其在大鼠肾脏缺血再灌注损伤中的保护作用及其机制。方法：制备并检测HBSP的纯度及分子量。200~220g SD大鼠随机分为假手术组、缺血组、HBSP组。阻断左侧肾蒂45min后切除右侧肾脏模型。术中及术后每6h给予HBSP,检测术后48h肾功能及炎症细胞因子肿瘤坏死因子-α（TNF-α）,观察病理学改变,采用末端转移酶标记技术（TUNEL）法检测肾小管上皮细胞凋亡,并检测肾组织核转录因子-κB（NF-κB）活性。结果：HBSP纯度达98%,分子量与理论分子量吻合,HBSP组术后48h肾功能好于缺血组,组织中TNF-α水平及NF-κB活性显著低于缺血组（P〈0.05）,HE切片组织损伤减轻,TUNEL染色示HBSP组阳性细胞数显著少于缺血组（P〈0.05）。结论：由促红细胞生成素成功制备线性多肽HBSP,其通过减轻炎性反应及抑制肾小管细胞凋亡,保护肾脏缺血再灌注损伤大鼠的肾功能。

Objective：To prepare and identify the helix B-surface peptide（HBSP）,a erythropoietin（EPO） derivative peptide,and investigate its protective effect on renal ischemia-reperfusion（IR） injury.Methods：HBSP was synthesized and identified.In order to investigate the role of renal protection,48 male Sprague-Dawley rats（200-220 g） were divided into three groups,Group Sham,Group IR and Group HBSP.IR model was made by 45 minutes of left renal ischemia with non-traumatic vascular clamps and right nephrectomy after the clamp was released.Serum creatinine,urea nitrogen and renal pathology were detected as well as the expression of tumor necrosis factor alpha（TNF-α）,nuclear factor-kappa B（NF-κB）.Apoptosis of tubular epithelium was observed by the terminal deoxynucleotidyl transferase-midiated dUTP-biotin nick end-labeling（TUNEL） assay.Results： The product purity was 98.2%,and the molecular weight was the same as we hypothesized before.Both serum creatinine and urea nitrogen were decreased in Group HBSP,accompanied with significantly mild histological damage and less TUNEL-positive cells.The expression level of TNF-α and NF-κB in Group HBSP was also decreased.Conclusions： HBSP is successfully prepared and indentified.HBSP attenuates renal ischemia reperfusion injury.The protective effect is mediated by its inhibition of inflamation and tubular epithelium apoptosis.